Cyclic uniaxial cell stretching in tissue culture using a LEGO®-based mechanical stretcher and a polydimethylsiloxane stretchable vessel.
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ABSTRACT:
SUBMITTER: Boulter E
PROVIDER: S-EPMC8055712 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:It has been previously demonstrated that uniaxial cyclic stretching (UCS) induces differentiation of mesenchymal stem cells (MSCs) into osteoblasts in vitro. It is also known that interactions between cells and external forces occur at various aspects including cell-matrix, cytoskeleton, nucleus membrane, and chromatin. However, changes in chromatin landscape during this process are still not clear. The present study was aimed to determine changes of chromatin accessibility under cyclic stretch. The influence of cyclic stretching on the morphology, proliferation, and differentiation of hMSCs was characterized. Changes of open chromatin sites were determined by assay for transposase accessible chromatin with high-throughput sequencing (ATAC-seq). Our results showed that UCS induced cell reorientation and actin stress fibers realignment, and in turn caused nuclear reorientation and deformation. Compared with unstrained group, the expression of osteogenic and chondrogenic marker genes were the highest in group of 1 Hz + 8% strain; this condition also led to lower cell proliferation rate. Furthermore, there were 2022 gene loci with upregulated chromatin accessibility in 1 Hz + 8% groups based on the analysis of chromatin accessibility. These genes are associated with regulation of cell morphogenesis, cell-substrate adhesion, and ossification. Signaling pathways involved in osteogenic differentiation were found in up-regulated GO biological processes. These findings demonstrated that UCS increased the openness of gene loci associated with regulation of cell morphogenesis and osteogenesis as well as the corresponding transcription activities. Moreover, the findings also connect the changes in chromatin accessibility with cell reorientation, nuclear reorientation, and deformation. Our study may provide reference for directed differentiation of stem cells induced by mechanical microenvironments.
| S-EPMC8294092 | biostudies-literature
Project description:Mechanical cues including stretch, compression, and shear stress play a critical role in regulating the behavior of many cell types, particularly those that experience substantial mechanical stress within tissues. Devices that impart mechanical stimulation to cells in vitro have been instrumental in helping to develop a better understanding of how cells respond to mechanical forces. However, these devices often have constraints, such as cost and limited functional capabilities, that restrict their use in research or educational environments. Here, we describe a low-cost method to fabricate a uniaxial cell stretcher that would enable widespread use and facilitate engineering design and mechanobiology education for undergraduate students. The device is capable of producing consistent and reliable strain profiles through the use of a servomotor, gear, and gear rack system. The servomotor can be programmed to output various waveforms at specific frequencies and stretch amplitudes by controlling the degree of rotation, speed, and acceleration of the servogear. In addition, the stretchable membranes are easy to fabricate and can be customized, allowing for greater flexibility in culture well size. We used the custom-built stretching device to uniaxially strain macrophages and cardiomyocytes, and found that both cell types displayed functional and cell shape changes that were consistent with the previous studies using commercially available systems. Overall, this uniaxial cell stretcher provides a more cost-effective alternative to study the effects of mechanical stretch on cells, and can therefore, be widely used in research and educational environments to broaden the study and pedagogy of cell mechanobiology.
| S-EPMC6056193 | biostudies-literature
Project description:ObjectiveThe retina is subjected to tractional forces in various conditions. As the predominant glial element in the retina, Müller cells are active players in all forms of retinal injury and disease. In this study, we aim to identify patterns of gene expression changes induced by cyclic mechanical stretching in Müller cells.MethodsRat Müller cells were seeded onto flexible bottom culture plates and subjected to a cyclic stretching regimen of 15% equibiaxial stretching for 1 and 24 h. RNA was extracted and amplified, labeled, and hybridized to rat genome microarrays. The expression profiles were analyzed using GeneSpring software, and gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to select, annotate, and visualize genes by function and pathway. The selected genes of interest were further validated by Quantitative Real-time PCR (qPCR).ResultsMicroarray data analysis showed that at 1 and 24 h, the expression of 532 and 991 genes in the Müller cells significantly (t-test, p<0.05) differed between the mechanically stretched and unstretched groups. Of these genes, 56 genes at 1 h and 62 genes at 24 h showed more than a twofold change in expression. Several genes related to response to stimulus (e.g., Egr2, IL6), cell proliferation (e.g., Areg, Atf3), tissue remodeling (e.g., PVR, Loxl2), and vasculogenesis (e.g., Epha2, Nrn1) were selected and validated by qPCR. KEGG pathway analysis showed significant changes in MAPK signaling at both time points.ConclusionsCyclic mechanical strain induces extensive changes in the gene expression in Müller cells through multiple molecular pathways. These results indicate the complex mechanoresponsive nature of Müller cells, and they provide novel insights into possible molecular mechanisms that would account for many retinal diseases in which the retina is often subjected to mechanical forces, such as pathological myopia and proliferative vitreoretinopathy.
| S-EPMC3664568 | biostudies-literature
Project description:This paper demonstrates a fabrication technology of Ag wrinkled electrodes with application in highly stretchable wireless sensors. Ag wrinkled thin films that were formed by vacuum deposition on top of pre-strained and relaxed polydimethylsiloxane (PDMS) substrates which have been treated using an O2 plasma and a surface chemical functionalization process can reach a strain limit up to 200%, while surface adhesion area can reach 95%. The electrical characteristics of components such as resistors, inductors and capacitors made from such Ag conductors have remained stable under stretching exhibiting low temperature and humidity coefficients. This technology was then demonstrated for wireless wearable electronics using compatible processing with established micro/nano fabrication technology.
| S-EPMC4653735 | biostudies-other
Project description:We present a pinned insect manipulator (IMp) constructed of LEGO® building bricks with two axes of movement and two axes of rotation. In addition we present three variants of the IMp to emphasise the modular design, which facilitates resizing to meet the full range of pinned insect specimens, is fully customizable, collapsible, affordable and does not require specialist tools or knowledge to assemble.
| S-EPMC4319104 | biostudies-literature
Project description:Cells respond to mechanical cues from their environment through a process of mechanosensing and mechanotransduction. Cell stretching devices are important tools to study the molecular pathways responsible for cellular responses to mechanobiological processes. We describe the development and testing of a uniaxial cell stretcher that has applications for microscopic as well as biochemical analyses. By combining simple fabrication techniques with adjustable control parameters, the stretcher is designed to fit a variety of experimental needs. The stretcher can be used for static and cyclic stretching. As a proof of principle, we visualize stretch induced deformation of cell nuclei via incremental static stretch, and changes in IEX1 expression via cyclic stretching. This stretcher is easily modified to meet experimental needs, inexpensive to build, and should be readily accessible for most laboratories with access to 3D printing.
| S-EPMC7888744 | biostudies-literature
Project description:Mechanical cues from the extracellular microenvironment play a central role in regulating the structure, function and fate of living cells. Nevertheless, the precise nature of the mechanisms and processes underlying this crucial cellular mechanosensitivity remains a fundamental open problem. Here we provide a novel framework for addressing cellular sensitivity and response to external forces by experimentally and theoretically studying one of its most striking manifestations--cell reorientation to a uniform angle in response to cyclic stretching of the underlying substrate. We first show that existing approaches are incompatible with our extensive measurements of cell reorientation. We then propose a fundamentally new theory that shows that dissipative relaxation of the cell's passively-stored, two-dimensional, elastic energy to its minimum actively drives the reorientation process. Our theory is in excellent quantitative agreement with the complete temporal reorientation dynamics of individual cells measured over a wide range of experimental conditions, thus elucidating a basic aspect of mechanosensitivity.
| S-EPMC4066201 | biostudies-literature
Project description:Purpose Effective communication skills are a critical quality and skill that is highly sought after for surgeons which largely impacts patient outcomes. Residency programs design their interview processes to select the best candidates. LEGO®-based activities have been frequently used to enhance communication skills and team building. This study investigates the effectiveness and reliability of a novel LEGO®-based communication assessment in interviews for surgical residencies and the feasibility of implementing it in a virtual setting. Methods This study conducted a retrospective analysis of a LEGO®-based communication assessment at the program’s 2020/2021 residency interviews. Each applicant was assessed on a different model. The total scores were analyzed for consistency among raters and correlated to faculty interviews. Furthermore, the impact of the assessment structure, scoring criteria, and range of models’ difficulties on the total scores were explored. Results A total of 54 categorical and 55 preliminary applicants interviewed on 2 days. The assessment on different models and had no impact on applicants’ total scores for either categorical and preliminary groups (p = 0.791 and 0.709, respectively). The communication components of the assessment showed high consistency between the raters. The two applicant groups displayed a statistically significant difference (p = 0.004) in the communication evaluation and model accuracy components. Total scores did not correlate with the faculty interviews of standardized questions in either group. Conclusion This novel LEGO®-based communication assessment showed high reliability and promising results as a tool to assess communication and problem solving for residency interviews that can be readily implemented in a virtual setting. Supplementary Information The online version contains supplementary material available at 10.1007/s44186-022-00021-4.
| S-EPMC9171471 | biostudies-literature
Project description:Graphene-based optical sensing devices have been widely studied for their broad band absorption, high carrier mobility, and mechanical flexibility. Due to graphene's weak light absorption, studies on graphene-based optical sensing thus far have focused on hybrid heterostructure devices to enhance photo-absorption. Such hybrid devices need a complicated integration process and lead to deteriorating carrier mobility as a result of heterogeneous interfaces. Rippled or wrinkled graphene has been studied in electronic and optoelectronic devices. However, concrete demonstrations of the impact of the morphology of nanofilms (e.g., graphite and graphene) associated with light absorption in optical sensing devices have not been fully examined. This study explored the optical sensing potential of a graphite nanofilm surface with ripples induced by a stretchable polydimethylsiloxane (PDMS) supporting layer under different stretch:release ratios and then transferred onto silicon, both under experimental conditions and via simulation. The optical sensing potential of the rippled graphite nanofilm was significantly enhanced (260 mA/W at the stretch-release state of 30%), as compared to the pristine graphite/PDMS (20 mA/W at the stretch-release state of 0%) under laser illumination at a wavelength of 532 nm. In addition, the results of our simulated computation also confirmed the improved light absorption of rippled graphite nanofilm surface-based optical sensing devices, which was comparable with the results found in the experiment.
| S-EPMC8619932 | biostudies-literature
Project description:To study the effects of mechanical stretching on fibroblasts, two cell lines, HFF and KF, were subject to 50% uniaxial mechanical stretching for 6, 12, and 18 hours, followed by RNA-seq.
2024-06-05 | GSE262112 | GEO